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Characterization of Two Kinases Involved in Thiamine Pyrophosphate and Pyridoxal Phosphate Biosynthesis in Bacillus subtilis: 4-Amino-5-Hydroxymethyl-2-Methylpyrimidine Kinase and Pyridoxal Kinase. Joo-Heon Park, 2004.Two Bacillus subtilis genes encoding two proteins [currently annotated ThiD and YjbV] were overexpressed and characterized.YjbV has 4-amino-5-hydroxymethyl-2-methylpyrimidine and 4-amino-5-hydroxymethyl-2-methylpyrimidinepyrophosphate kinase activity and should be reannotated ThiD,and B . subtilis ThiD has pyridoxine, pyridoxal, and pyridoxaminekinase activity and should be reannotated PdxK. Possible Mechanism of Miltefosine-Mediated Death of Leishmania donovani. Navin K. Verma, 2004.Miltefosine causes leishmanial death, but the possible mechanism(s) of action is not known . The mode of action of miltefosine was investigated in vitro in Leishmania donovani promastigotes as well as in extra- and intracellular amastigotes . Here, we demonstrate that miltefosine induces apoptosis-like death in L . donovani based on observed phenomena such as nuclear DNA condensation, DNA fragmentation with accompanying ladder formation, and in situ labeling of DNA fragments by the terminal deoxyribonucleotidyltransferase-mediated dUTP-biotin nick end labeling method . Understanding of miltefosine-mediated death will facilitate the design of new therapeutic strategies against Leishmania parasites . Formation of dTDP-Rhamnose Is Essential for Growth of Mycobacteria. Yufang Ma, 2002.It was determined that the dTDP-rhamnose synthesis gene, rmlD, could be inactivated in Mycobacterium smegmatis only in the presence of a rescue plasmid carrying functional rmlD. Hence, dTDP-rhamnose biosynthesis is essential for the growth of mycobacteria and the targeting of dTDP-rhamnose synthesis for new tuberculosis drugs is supported . Metabolic Flux in Both the Purine Mononucleotide and Histidine Biosynthetic Pathways Can Influence Synthesis of the Hydroxymethyl Pyrimidine Moiety of Thiamine in Salmonella enterica. Shara Allen, 2002.Together, the biosyntheses of histidine, purines, and thiamine pyrophosphate (TPP) contain examples of convergent, divergent, and regulatory pathway integration . Mutations in two purine biosynthetic genes (purI and purH) affect TPP biosynthesis due to flux through the purine and histidine pathways . The molecular genetic characterization of purI mutants and their respective pseudorevertants resulted in the conclusion that <1% of the wild-type activity of the PurI enzyme was sufficient for thiamine but not for purine synthesis . The respective pseudorevertants were found to be informational suppressors . In addition, it was shown that accumulation of the purine intermediate aminoimidazole carboxamide ribotide inhibits thiamine synthesis, specifically affecting the conversion of aminoimidazole ribotide to hydroxymethyl pyrimidine .
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